A Putative Mechanism for Magnetoreception by Electromagnetic Induction in the Pigeon Inner Ear
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Report A Putative Mechanism for Magnetoreception by Electromagnetic Induction in the Pigeon Inner Ear Graphical Abstract Authors Simon Nimpf, Gregory Charles Nordmann, Daniel Kagerbauer, ..., Martin Colombini, Matthew J. Mason, David Anthony Keays Correspondence [email protected] In Brief Nimpf, Nordmann et al. confirm that magnetic stimuli result in neuronal activity in the vestibular nuclei of pigeons. Hypothesizing that this is attributable to electromagnetic induction within semicircular canals of the inner ear, they demonstrate the presence of known electrosensory molecules in vestibular hair cells. Highlights d Magnetic stimuli activate neurons in the caudal vestibular nuclei d Magnetic stimuli induce a voltage in a model of a semicircular canal d Electroreceptive molecules are expressed in vestibular hair cells d We postulate that pigeons detect magnetic fields by electromagnetic induction Nimpf et al., 2019, Current Biology 29, 4052–4059 December 2, 2019 ª 2019 The Author(s). Published by Elsevier Ltd. https://doi.org/10.1016/j.cub.2019.09.048 Current Biology Report A Putative Mechanism for Magnetoreception by Electromagnetic Induction in the Pigeon Inner Ear Simon Nimpf,1,5 Gregory Charles Nordmann,1,5 Daniel Kagerbauer,2 Erich Pascal Malkemper,1 Lukas Landler,1 Artemis Papadaki-Anastasopoulou,1 Lyubov Ushakova,1 Andrea Wenninger-Weinzierl,1 Maria Novatchkova,1 Peter Vincent,3 Thomas Lendl,1 Martin Colombini,1 Matthew J. Mason,4 and David Anthony Keays1,6,* 1Research Institute of Molecular Pathology (IMP), Vienna Biocenter (VBC), Campus-Vienna-Biocenter 1, 1030 Vienna, Austria 2Atominstitut, TU Wien, Stadionallee 2, 1020 Vienna, Austria 3Department of Neurology, Yale University School of Medicine, 333 Cedar Street, New Haven, CT 06520, USA 4Department of Physiology, Development and Neuroscience, University of Cambridge, Downing Street, Cambridge CB2 3EG, UK 5These authors contributed equally 6Lead Contact *Correspondence: [email protected] https://doi.org/10.1016/j.cub.2019.09.048 SUMMARY activation in the vestibular nuclei of pigeons [5]. To perform this experiment within the laboratory environment, we built a A diverse array of vertebrate species employs the room constructed of mu metal surrounded by an aluminum Earth’s magnetic field to assist navigation. Despite Faraday cage to shield against static and oscillating magnetic compelling behavioral evidence that a magnetic fields (Figure 1A). This setup allowed us to perform experiments sense exists, the location of the primary sensory cells in a controlled, magnetically clean environment [7]. Magnetic and the underlying molecular mechanisms remain un- fields were generated using a double-wrapped, custom-built known [1]. To date, most research has focused on a 3D Helmholtz coil system situated in the center of the shielded room (Figure 1B). To reduce movement during the experiments, light-dependent radical-pair-based concept and a birds were head fixed using a surgically implanted plastic head system that is proposed to rely on biogenic magnetite post, and the body was immobilized using a 3D printed harness (Fe3O4)[2, 3]. Here, we explore an overlooked hypoth- (Figure 1C). We applied the same stimulus as Wu and Dickman, esis that predicts that animals detect magnetic fields exposing adult pigeons to a 150-mT, rotating magnetic field (n = by electromagnetic induction within the semicircular 22) or to a zero magnetic field (n = 23) for 72 min (Figures 1D–1F) canals of the inner ear [4]. Employing an assay that re- [5]. We performed this experiment both in darkness (n = 30) and lies on the neuronal activity marker C-FOS, we under broad-spectrum white light (n = 15). Birds were then confirm that magnetic exposure results in activation perfused, the brains were sliced, and matched sections contain- of the caudal vestibular nuclei in pigeons that is inde- ing the vestibular nuclei (3 sections per bird) were stained with pendent of light [5]. We show experimentally and by sera against the neuronal activity marker C-FOS. To minimize physical calculations that magnetic stimulation can variation, all staining was performed simultaneously, all slides were scanned with the same exposure settings, and C-FOS- induce electric fields in the pigeon semicircular canals positive neurons were counted using a machine-learning-based that are within the physiological range of known elec- algorithm. Using established anatomical coordinates, we troreceptive systems. Drawing on this finding, we segmented the medial vestibular nuclei (VeM) and compared report the presence of a splice isoform of a voltage- the density of C-FOS-positive cells of the experimental groups gated calcium channel (CaV1.3) in the pigeon inner [8]. We observed an increase in the density of C-FOS-positive ear that has been shown to mediate electroreception cells in both the light and dark when exposing birds to magnetic in skates and sharks [6]. We propose that pigeons fields. In the light, the average density in controls was 35.32 ± detect magnetic fields by electromagnetic induction 10.20 cells/mm2 (n = 8; mean ± SD) whereas with the magnetic 2 within the semicircular canals that is dependent on treatment it was 44.38 ± 5.72 cells/mm (n = 7; mean ± SD). In the dark, the average density for control birds was 34.70 ± the presence of apically located voltage-gated cation 2 channels in a population of electrosensory hair cells. 10.82 cells/mm (n = 15; mean ± SD) whereas with the magnetic treatment it was 50.52 ± 27.34 cells/mm2 (n = 15; mean ± SD). An application of a two-way ANOVA revealed a significant RESULTS effect of the magnetic treatment but no interaction between magnetic treatment and lighting conditions (two-way ANOVA; Magnetically Induced Activation in the Pigeon magnetic: p = 0.0176, F = 6.125; magnetic by light: p = 0.5675, Vestibular Brainstem F = 0.332) (Figures 2A and 2B; Table S1). To explore this in We set out to replicate a previous study conducted by Wu and more detail, we employed our spot-detection algorithm coupled Dickman, who reported that magnetic stimuli induce neuronal to an elastic registration to generate heatmaps showing regional 4052 Current Biology 29, 4052–4059, December 2, 2019 ª 2019 The Author(s). Published by Elsevier Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). Figure 1. Experimental Setup for the Magnetic Activation Study (A) The magnetic activation experiments were performed inside a magnetically shielded room consisting of a Faraday cage to shield from broadband anthro- pogenic magnetic noise and a mu-metal cage to shield from the geomagnetic field. (B and C) Pigeons were placed in the center of a double-wrapped 3D Helmholtz coil system (B) located in the magnetically shielded room and immobilized by head fixation and a molded body harness (C). (D–F) Schematic illustration of the magnetic stimulus used in this study. The magnetic field vector was rotated 360 within a plane by 6 steps every 2 s (D). After each rotation the plane was shifted by 15 (E) and the procedure was repeated until a full rotation around each axis was achieved (12 planes per x, y, and z axis, respectively) (F). differences in the density of C-FOS-positive cells. This revealed nanovoltmeter (forming a loop with a diameter of 21 cm), placed an enrichment of activated neurons in the dorsomedial part of in the center of our magnetic coil system, and exposed to the the VeM in animals exposed to magnetic stimuli (Figure 2C). Pre- same rotating magnetic stimulus applied to our birds (i.e., vious tracing experiments have shown that the dorsomedial VeM 150 mT rotating 360 with 6-step changes every 2 s). We is innervated by projections from both the semicircular canals observed discrete voltage spikes when the magnetic field stim- and the otolith organs located in the inner ear [9]. ulus was presented and with each stepwise change (Figure 3B). In accordance with Faraday’s law of electromagnetic induction, A Model for Electromagnetic Induction in the Pigeon we observed the maximum induced voltage (15.6 mV) when the Inner Ear magnetic field vector was perpendicular (90) to the plane of As we did not observe an interaction between the presence of the canal and lowest (1.6 mV) when the vector was parallel to light and magnetically induced neuronal activation, our results the plane of the canal (0)(Figure 3B). In contrast, when present- are consistent with a magnetic sensory system based on either ing the control stimulus (with the current running antiparallel magnetite or electromagnetic induction [10]. We have previously through the double-wrapped coils), we did not observe these reported the discovery of an iron-rich organelle in both vestibular characteristic voltage spikes (Figure 3C). Nor did we observe and cochlear hair cells that is associated with vesicular struc- voltage spikes when removing the model of the semicircular ca- tures, but because it is primarily composed of ferrihydrite it lacks nal from the circuit, demonstrating that induction does not occur the magnetic properties to function as the hypothesized torque- in the connecting shielded wires (Figure 3D). based magnetoreceptor [11–13]. Moreover, a systematic screen Next, we measured the dimensions of the semicircular canals for magnetite in the pigeon lagena using synchrotron-based in pigeons drawing on previously generated computed tomogra- X-ray fluorescence microscopy and electron microscopy has phy (CT) scans (n = 3 birds) [18](Figure S1A). This revealed that failed to identify extra- or intracellular magnetite crystals [14]. In the mean loop diameter of the posterior canal was 4.20 ± light of these findings, we focused on electromagnetic induction 0.01 mm, the anterior canal was 6.37 ± 0.05 mm, and the lateral [15]. First proposed by Camille Viguier in 1882, this hypothesis canal was 5.18 ± 0.15 mm (mean ± SD). Drawing on these mea- predicts that as a terrestrial animal moves through the Earth’s surements and the voltage induced in our 21-cm-diameter static magnetic field a voltage is induced within the conductive model, we were able to estimate the electric field generated endolymph of the semicircular canals [4, 16].